Objective To construct a chimeric infectious clone of the fatal virulent strain SDLY 107, containing the gene fragments encoding 2A and 3B proteins of the mild virulent strain SDLY 1, and to establish a reverse genetic system platform for further investigation on virulence of enterovirus 71 strains. Methods The overlap PCR analysis was performed to obtain the gene fragments encoding 2A and 3B pro-teins of the mild virulent strain SDLY 1.The obtained gene fragments were digested and then cloned into a plasmid pMD19-T containing the full-length gene of SDLY 107 strain by using gene replacement strategy. The recombinant RNA was transfected into Vero cells for the preparation of recombinant virus particles.Sev-eral assays including the PCR, indirect immunofluorescence ( IFA) , Western blot and sequencing were per-formed for virus identification.Virus titers were measured by 50%cell culture infective dose ( CCID50 ) and plaque assay.Results The infectious clones of SDLY 107-2A-1 and SDLY 107-3B-1 chimeric virus strains were constructed successfully.Typical cytopathic effect was observed in Vero cells after viral transfection. Identification of the rescued viruses by PCR, IFA, Western blot and sequencing further confirmed the suc-cessful construction of infectious virus strains.The virus titers of SDLY 107-2A-1 and SDLY 107-3B-1 strains detected by CCID50 and plaque assay were 1.25 ×105 PFU/ml and 0.7 ×105 PFU/ml, respectively. Conclusion The chimeric viruses SDLY 107-2A-1 and SDLY 107-3B-1 were rescued successfully, causing cytopathic effects similar to those by using the parental virus strain SDLY 107.This study might pave the way for further investigation on in vitro and in vivo virulence of enterovirus 71 strains.

OBJECTIVETo study the proliferation and differentiation of neural stem cells in the subventricular zone (SVZ) in neonatal rats after bilateral common arteries occlusion.

METHODSNinety-six 3-day-old Sparuge-Dawley rats were randomly divided into two groups: ischemia and control. Rats in the ischemia group were subjected to bilateral common arteries occlusion and the rats in the control group were sham-operated. All rats were administrated with 5-bromodeoxyuridine (BrdU) (50 mg/kg) via intraperitoneal injection. Rats were sacrificed and their brains were removed 1, 4, 7, 10, 14 and 35 days after ischemia. Using brain paraffin sections and immunofluorescence assays, the number of newborn cells in the SVZ was counted. Newborn neural stem cells and oligodendrocytes in the SVZ were observed, and then double marked with BrdU and nestin or osmium tetroxide (O4).

RESULTSThe number of BrdU+ cells (neural stem cells) in the SVZ in the ischemia group was greater than in the control group 4, 7, 10 and 14 days after ischemia, and reached a peak at 4 days after ischemia (253.1+/- 49.3 vs 133.5+/- 17.7; P< 0.01). By 35 days after ischemia, the number of BrdU+/O4+ cells (oligodendrocytes) in the corpus callosum (56.0+/- 7.2 vs 17.0+/- 6.4; P< 0.01), the septal nuclei (45.0+/- 11.9 vs 20.5+/- 5.0; P< 0.01), the striatum (34.5+/- 4.2 vs 14.5+/- 5.8; P< 0.01) and the olfactory bulb (46.5+/- 6.6 vs 23.5+/- 8.4; P< 0.01) in the ischemia group increased significantly as compared to the control group (P< 0.01).

CONCLUSIONSBrain ischemia can activate the proliferation of neural stem cells in the SVZ and promote neural stem cells differentiation into oligodendrocytes. The immature brain may have the capacity for self-repair after ischemic brain injury.

Animals ; Animals, Newborn ; Brain Ischemia ; physiopathology ; therapy ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cerebral Ventricles ; physiopathology ; Female ; Male ; Neurogenesis ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation

Objective To Design a medical information system to meet the requirements for informatized medical support by the field medical support institution. Methods J2EE architecture and Java technology were applied to the design of the system, and star topology was used for network deployment. Results The system shared casualty treatment data with servicemen electronic tag and medical commanding information with logistics integrated command and control platform,so that the requirements for medical information were met in the institution.Conclusion The system improves the field medical support institution in medical support ability,treatment rate and efficacy.

Objective To investigate the effects of in situ venous arterialization on extensive artery obliterans occlusion of the lower extremity. Methods Lumbar sympathetic ganglionectomy and one stage in situ arterialization of the great saphemous vein were performed in 104 ischemic limbs of 88 patients with extensive arterial occlusion. Results Eighty-two of 104 limbs were followed-up from 6 months to over 6 years. The intermittent claudication, night pain improved in all cases, with satisfactory wound healing and no swelling of the lower limbs. Conclusions Arterial blood flow through venous conduit improves and reconstructs the blood circulation of the ischemic limbs.

OBJECTIVETo analyze the tendency in the incidence of malignant tumors in nutrition-intervened population of Lin County.

METHODSSubjects who had been enrolled in the General Population Trial, Lin County Nutrition Intervention Trial were followed up and the data of malignant tumors were collected with collaboration of National Cancer Institute/National Institute of Health. Incidences of malignant tumors during and after nutrition intervention were calculated and compared.

RESULTSEsophageal cancer, cardiac cancer, and stomach cancer were constantly the three leading cancers, accounting for 84.06% and 83.74% of the malignant tumors in men and women, respectively. The incidences of lung cancer and liver cancer in men showed increasing tendencies, while similar tendencies were found in those of stomach cancer, colorectal cancer, and liver cancer in women. With the prolonging of the follow-up, the incidences of upper digestive tract cancers declined (except stomach cancer in women).

CONCLUSIONSThe upper digestive tract cancers remain the leading malignant tumors, although their incidences may decline with longer-term follow-up. Prevention and control of tumors in Lin County should still be focused on the esophageal cancer, cardiac cancer, and stomach cancer, as well as lung cancer and liver cancer.

Adult ; Aged ; China ; epidemiology ; Diet ; Feeding Behavior ; Female ; Follow-Up Studies ; Humans ; Incidence ; Male ; Middle Aged ; Neoplasms ; epidemiology ; Prospective Studies ; Risk Factors

BACKGROUNDLaparoendoscopic single-site surgery (LESS) approaches have been reported for treating various kidney and pelvic procedures, and are feasible and effective in selected patients. In this study, we aimed to present the initial experience and evaluate the efficacy of laparoscopic radical prostatectomy performed through a single incision using a multichannel port.

METHODSBetween July 2010 and April 2011, six patients diagnosed with early stage prostate cancer underwent LESS radical prostatectomy (RP) in our institute. A multichannel port was inserted transperitoneally through a 2-cm umbilical incision. Specially articulating and flexible laparoscopic were used. Some technical tricks and points were applied during the operation to overcome the drawbacks and reduce the difficulties of this approach. Two continuous urethrovesical sutures in both sides were performed to complete both lateral aspects of anastomosis. The two ends of the suture threads were fixed by double Lapro-Clips, instead of the difficult knot-tying.

RESULTSTotal operative time was (265 ± 43) minutes. Mean blood loss was (230 ± 65) ml. All cases were completed successfully, without conversion to open surgery or adding additional abdomen ports. No patient required a blood transfusion and no intraoperative complications occurred. The Foley catheter was removed at the 14th day (range 12th - 16th) after surgery. At the 12th week of follow-up, all patients had an undetectable prostate-specific antigen level. Two patients used 2 or 1 pad for continence daily; other patients had achieved good continence.

CONCLUSIONIn selected cases, LESS-RP is feasible and effective; these technic points and the flexible-articulating instruments are helpful to reduce the operation difficulties.

Aged ; Humans ; Laparoscopy ; methods ; Male ; Prostatectomy ; methods

OBJECTIVETo explore effect of interleukin 10 (IL-10) gene-modified bone marrow-derived liver stem cells (BDLSCs) transplantation on hepatic inflammatory response and liver regeneration in rats with liver fibrosis.

METHODS50 female Wistar rats were randomly divided into 4 groups: (1) control group: 10 rats were subcutaneously injected with olive oil for 8 weeks; (2) fibrosis groups: 16 rats were subcutaneously injected with carbon tetrachloride (CCl4) for 8 weeks to induce liver fibrosis; (3) BDLSC group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) BDLSC at week 4; (4) BDLSC/IL-10 group: 12 rats were subcutaneously injected with CCl4 for 8 weeks, and were transplanted with 2 x 10(5) IL-10 gene-modified BDLSC at week 4. IL-10 and tumor necrosis factor alpha (TNFa) in liver tissues were detected by ELISA. HE stained liver tissues were observed under light microscope. The expression of hepatocyte growth factor (HGF) was quantified by real-time RT-PCR, and the expression of proliferating cell nuclear antigen (PCNA) was determined by immunohistochemistry.

RESULTSThe ratio of IL-10/TNFa in fibrosis group (0.05+/-0.01) was lower than that in control group (0.26+/-0.04) (P < 0.01). Transplantation of untreated BDLSCs did not improve the ratio (P > 0.05), however, transplantation of IL-10 modified BDLSCs improved the ratio significantly (P < 0.01). Severe inflammatory response and fibrosis were observed in fibrosis group. Inflammatory response was alleviated to some extent in the BDLSC group, and the histopathology of BDLSC/IL-10 group was not significantly different from that of the control group. Compared to the control group, the expression of HGF mRNA and PCNA protein was increased in the fibrosis group (P < 0.01). The expression of HGF and PCNA was further increased by BDLSCs or IL-10 modified BDLSCs transplantation. Compared to BDLSCs, IL-10 gene-modified BDLSCs were more potent to induce the expression of HGF and PCNA.

CONCLUSIONTransplantation of IL-10 gene-modified BDLSCs can alleviate hepatic inflammatory response and promote liver regeneration in hepatic fibrosis rats.

Adenoviridae ; genetics ; Animals ; Bone Marrow Cells ; cytology ; Cell Proliferation ; Disease Models, Animal ; Female ; Genetic Therapy ; methods ; Hepatocyte Growth Factor ; genetics ; metabolism ; Immunohistochemistry ; Interleukin-10 ; genetics ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; genetics ; pathology ; therapy ; Liver Regeneration ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Transplantation ; methods ; Transduction, Genetic ; Tumor Necrosis Factor-alpha ; metabolism

AIM:To investigate the effects of simvastatin on the expression of Toll-like receptor 2 ( TLR-2 ) , interferon-γ(IFN-γ) and monocyte chemoattractant protein-1 (MCP-1) in lung tissues of mice with mouse cytomegalovirus ( MCMV) pneumonia and to explore the possible mechanism .METHODS:Male BALB/c mice (6~8 weeks old, n=40) were randomly divided into 5 groups: normal control (NC) group, MCMV infection group, simvastatin group 1 (SMV1 group), simvastatin group 2 (SMV2 group), and simvastatin group 3 (SMV3 group).The mice in SMV1, SMV2 and SMV3 groups were gavaged with simvastatin (50 mg· kg-1 · d-1 for 7 d) 7 d before, on the same day of and 3 d after in-traperitoneal injection of MCMV , while the mice in normal control group and MCMV infection group were gavaged with the same volume of normal saline .HE staining was used to observe the pathological changes of lung tissues in mice .Total tis-sue protein was extracted from the lung homogenates to detect the expression of TLR-2 by Western blot and immunohisto-chemical staining .Real-time PCR was used to analyse the content of MCMV DNA .The levels of IFN-γand MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA).RESULTS:Compared with NC group, the pathological chan-ges of the lung tissues of the mice in MCMV group showed alveolar interstitial edema , alveolar wall widening and a large number of inflammatory cells .The expression of TLR-2 in the lung tissues of the mice in model group was increased signifi-cantly.The content of MCMV DNA was increased , and the expression of IFN-γand MCP-1 was also increased significant-ly.Compared with the mice in MCMV group , the pathological changes of the lung tissues of simvastatin groups showed that the inflammatory cells were decreased .The expression of TLR-2 was down-regulated.The content of MCMV DNA was de-creased, and the levels of IFN-γand MCP-1 were also decreased significantly .At the same time, the expression of TLR-2 and the content of MCMV DNA in SMV1 group were less than those in SMV2 and SMV3 groups (P<0.05), and no statis-tically significant difference between SMV 2 and SMV3 groups was observed .CONCLUSION:Simvastatin down-regulates the TLR-2 signaling pathway , and reduces the expression of TLR-2 and replication of MCMV DNA , thus attenuating the pathological damage of the lung tissue .Early intervention with simvastatin plays an important role in preventing the infection of MCMV and reducing the inflammation .

Objective: To study the rule pattern of risk factor of stroke through data analysis of preliminary screening of large-scale stroke. Methods: The preliminary screening data of risk factors in stroke were gathered from the National Center for Stroke Screening and Prevention and Control. And the association rules mining was adopted in the research. The relevant risk factors of stroke in large-scale crowd were researched, and 0.1% was chose as minimum support and 10% were used as minimum confidence coefficient. The rule mode of stroke was mined by using above conditions. Results: In the nine risk factors of the table of preliminary screening, there was a weak correlation between "transient ischemic attack (TIA) in past", "hypertension", "family history of stroke" and stroke, respectively. And TIA, atrial fibrillation or valvular heart disease, family history of stroke, hypertension and diabetes were the most important risk factors. At the same time, the study found that age would become an important risk factor for stroke when patient was older than 60. Conclusions: In the 21 presumptive rule patterns with big probability, some of them has not received sufficient attention in past. And the rule pattern of risk factor of stroke provides a new diagnostic basis with value.

OBJECTIVE: To explore the distribution of inflammatory cells and positive expression of P-se- lectin glycoprotein ligand-1 (PSGL-1) in infant brainstem tissue from hand-foot-mouth disease related fatal brainstem encephalitis. METHODS: Twenty brainstem samples from infants suffered from brainstem en- cephalitis were collected as the experimental group. Ten brainstem samples from infants died of non- brain diseases and injuries were collected as the control group. The distribution of inflammatory cells and the expression of PSGL-1 in the two groups were examined by immunohistochemical method. The characteristics of the positive cells were observed. RESULTS: In brainstem tissue of the experimental group, there were sleeve infiltrations of inflammatory cells around the vessels and in the glial nodule. Microglia was the most and following was neutrophils around the vessels and in the glial nodule. There was a significant statistical difference among microglias, neutrophils and lymphocytes (P < 0.05). There was no sleeve infiltration in the control group. PSGL-1 protein was expressed widely in inflammatory cells in the experimental group, especially in the inflammatory cells around the vessels and in the glial nodule. But PSGL-1 positive staining could be observed significantly less in the control group comparing with the experimental group (P < 0.05). CONCLUSION Microglia is the main type of inflammatory cells involved in the progress of the fatal disease. Moreover, PSGL-1 could participate in the pathogenesis of hand-foot-mouth disease related fatal brainstem encephalitis.
Brain Stem/pathology* ; Encephalitis/pathology* ; Hand, Foot and Mouth Disease/pathology* ; Humans ; Infant ; Membrane Glycoproteins/metabolism* ; Microglia/pathology* ; Neutrophils/pathology*